Revision 4

#97747

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, W-S

Reactivity:
All

Sensitivity:
Transfected Only

MW (kDa):
130

Source/Isotype:
Mouse IgG2a

UniProt ID:
#A0A355UCT5

Entrez-Gene Id:
999999

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

PspCas13b (Prevotella sp. P5–125) (12F2-A10) Mouse Monoclonal Antibody recognizes transfected levels of total PspCas13b (Prevotella sp. P5–125) protein. This antibody may detect several non-specific bands of unknown origin in rat cell lines.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of PspCas13b (Prevotella sp. P5–125) protein.

Background

CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins) are RNA-guided nuclease effectors that are utilized for precise genome editing in mammalian systems (1). Cas13 proteins are members of the Class 2 CRISPR-Cas system. Class 2 CRISPR systems, such as the well characterized Cas9, rely on single-component effector proteins to mediate DNA/RNA interference rather than the multi-Cas protein complexes that belong to the Class 1 systems (2). Evolutionarily, CRISPR-Cas proteins act as RNA-guided antiviral and anti-plasmid that protect prokaryotes from phage-mediated infections. RNA-guided CRISPR-Cas proteins can act on both DNA and RNA. Cas13 proteins act as RNA-guided RNA nucleases, disrupting phage infection by RNA cleavage. The CRISPR-Cas system is being used for nuclease-based gene-editing. Since the Cas13 proteins act on RNA rather than DNA, Cas13 proteins, coupled with the appropriate guide RNA, can be used to target specific RNA substrates, potentially replacing or complementing RNA interference approaches that interfere with transcript levels (3). Several Cas13 bacterial orthologs have been characterized for CRISPR-mediated mammalian RNA editing, including LshCas13a/C2c2 and PspCas13b derived from Leptotrichia shahii and Prevotella sp. P5–125, respectively (4-6).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™

Cross-Reactivity Key

All: All Species Expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

Cell Signaling Technology Logo
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length Myc-tagged PspCas13b (PspCas13b-Myc; +), using PspCas13b (Prevotella sp. P5–125) (12F2-A10) Mouse mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: PspCas13b (<i>Prevotella sp. P5–125</i>) (12F2-A10) Mouse Monoclonal Antibody
Simple Western™ analysis of lysates (0.1 mg/mL) from 293T cells transfected with a construct expressing full-length Myc-tagged PspCas13b using PspCas13b (Prevotella sp. P5–125) (12F2-A10) Mouse mAb #97747. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western Blotting Image 1: PspCas13b (<i>Prevotella sp. P5–125</i>) (12F2-A10) Mouse Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.